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当前位置: 首页 > 文章 > Pseudomonas sp.W7产低温蛋白酶培养基及培养条件的优化 食品科学 2015,36 (19) 163-169
Position: Home > Articles > Optimization of Medium Components and Culture Conditions for Cold-Adapted Protease Produced by Pseudomonas sp.W7 FOOD SCIENCE 2015,36 (19) 163-169

Pseudomonas sp.W7产低温蛋白酶培养基及培养条件的优化

作  者：
康传红;田亚新;李秀凉;董义杰;原继红;王运来;韩晓云
单  位：
黑龙江大学化学化工与材料学院;黑龙江大学生命科学学院
关键词：
低温蛋白酶;假单胞菌W7;培养条件优化;响应面分析
摘  要：
采用Plackett-Burman法对影响假单胞菌(Pseudomonas sp.)W7产低温蛋白酶的因素进行显著性分析,筛选出对产酶影响显著的3个因素:葡萄糖、蛋白胨和MgSO_4;然后用最陡爬坡试验逼近最大产酶区域;应用Box-Behnken原理设计和响应面分析确定产酶培养基的最佳组合为:葡萄糖4.25 g/L、蛋白胨7.00 g/L、干酪素4.00 g/L、K2HPO_4 5.00 g/L、KH_2PO_4 1.00 g/L、CaCl_2 0.26 g/L、MgSO_4 0.14 g/L,低温蛋白酶的酶活力达到了183.9 U/mL,比优化前提高了21%。另外,对Pseudomonas sp.W7的产酶条件进行了优化,在单因素的基础上,利用正交试验设计,最终确定产酶的最优培养条件为:培养温度为20℃、初始pH值为7.0、接种量为3%、装液量为20%、摇床转速为100r/min,在最佳条件时测得的酶活力为193.2 U/mL。通过生长曲线和产酶曲线的测定,确定产酶的培养时间为48 h。
译  名：
Optimization of Medium Components and Culture Conditions for Cold-Adapted Protease Produced by Pseudomonas sp.W7
作  者：
KANG Chuanhong;TIAN Yaxin;LI Xiuliang;DONG Yijie;YUAN Jihong;WANG Yunlai;HAN Xiaoyun;School of Chemistry and Material Sciences,Heilongjiang University;School of Life Sciences,Heilongjiang University;
关键词：
cold-adapted protease;;Pseudomonas sp.W7;;optimization of fermentation conditions;;response surface analysis
摘  要：
Cold-adapted proteases have great superiority in the food,cosmetic,pharmaceutical and detergent industries due to high catalytic activity at low temperature and thermal instability.In the present study,the Plackett-Burman design was used to identify glucose,peptone and MgSO_4 as the most significant medium components affecting the production of cold-adapted protease by Pseudomonas sp.W7.Then the steepest ascent experiment was used to approach the optimal region of the three medium components.Box-Behnken design and response surface analysis were used to determine the best culture medium containing 4.25 g/L glucose,7.00 g/L peptone,4.00 g/L casein,5.00 g/L K_2HPO_4,1.00 g/L KH_2PO_4,0.26 g/L CaCl_2 and 0.14 g/L MgSO_4.After the optimization,the yield of the protease was increased by 21%to 183.9 U/mL.In addition,the optimal culture conditions for producing cold-adapted protease using the optimized medium were determined using combination of single factor experiments and orthogonal array design experiments as culture temperature of 20 ℃,initial medium pH of 7.0,inoculum amount of 3%,medium loading volume of 20%and rotation speed of 100 r/min.Under these conditions,the enzyme activity was 193.2 U/mL.Based on the growth curve and enzyme production curve,the optimal culture period was determined to be 48 h.

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